Factor VIII polymer conjugates

Abstract

The invention is a proteinaceous construct comprising a Factor VIII molecule which is conjugated to a water-soluble polymer via carbohydrate moieties of Factor VIII, and methods of preparing same.

1. A unit dose of a pharmaceutical composition effective to maintain a FVIII molecule at a level of at least 1% in a patient suffering from a bleeding disorder comprising (i) (a) a Factor VIII molecule; and (b) at least one water soluble polymer bound to said Factor VIII molecule, wherein said water soluble polymer is attached to the Factor VIII via one or more carbohydrate moieties located in the B domain of Factor VIII, wherein said molecule retains at least 50% of native FVIII activity, and (ii) a pharmaceutically acceptable excipient. 
     
     
       2. The unit dose of  claim 1 , wherein said composition is substantially free from albumin. 
     
     
       3. The unit dose of  claim 1 , wherein said water soluble polymer is attached to said carbohydrate moiety via a linker. 
     
     
       4. The unit dose of  claim 3 , wherein said linker is 4-[4-N-maleimidophenyl]butyric acid hydrazide (MBPH). 
     
     
       5. The composition of  claim 3 , wherein the linker is releasable or hydrolysable. 
     
     
       6. The unit dose of  claim 1 , wherein the pharmaceutical composition is housed in a vessel. 
     
     
       7. The unit dose of  claim 1 , wherein the pharmaceutical composition is housed in a syringe. 
     
     
       8. The unit dose of  claim 1 , in a form suited for injection. 
     
     
       9. The unit dose of  claim 8 , wherein the composition is in a powder form for reconstitution with a diluent prior to injection. 
     
     
       10. The unit dose of  claim 9 , reconstituted with a diluent. 
     
     
       11. The unit dose of  claim 1 , wherein the composition comprises a plurality of conjugates, each having at least one water soluble polymer bound to a native Factor VIII, said water soluble polymer attached to the modified Factor VIII via one or more carbohydrate moieties, said modified Factor VIII retaining at least 50% of native FVIII activity, and (ii) a pharmaceutically acceptable excipient. 
     
     
       12. The unit dose of  claim 1 , further comprising an unconjugated Factor VIII moiety. 
     
     
       13. The unit dose of  claim 1 , wherein each water-soluble polymer in the composition has a nominal average molecular weight in a range of from about greater than 5,000 Daltons to about 150,000 Daltons. 
     
     
       14. The unit dose of  claim 1 , wherein each water-soluble polymer in the composition is selected from the group consisting of polyethylene glycol (PEG), polysialic acid (PSA) and dextran. 
     
     
       15. The unit dose of  claim 14 , wherein each conjugate in the composition comprises the same water-soluble polymer. 
     
     
       16. The unit dose of  claim 15 , wherein the water-soluble polymer has a nominal average molecular weight in the range of from about 2,000 Daltons to about 150,000 Daltons. 
     
     
       17. The unit dose of  claim 16 , wherein the water-soluble polymer has a nominal average molecular weight in the range of from about 5,000 Daltons to about 50,000 Daltons. 
     
     
       18. The unit dose of  claim 15 , wherein each water-soluble polymer is linear. 
     
     
       19. The unit dose of  claim 15 , wherein each water-soluble polymer is branched. 
     
     
       20. The unit dose of  claim 15 , wherein the Factor VIII is recombinant Factor VIII. 
     
     
       21. The unit dose of  claim 15 , wherein the Factor VIII is plasmatic Factor VIII. 
     
     
       22. A kit comprising a unit dose of a pharmaceutical composition according to  claim 1  comprising (i) (a) a Factor VIII molecule; and (b) at least one water soluble polymer hound to said Factor VIII molecule, wherein said water soluble polymer is attached to the Factor VIII via one or more carbohydrate moieties located in the B domain of Factor VIII, wherein said molecule retains at least 50% of native FVIII activity, and (ii) a pharmaceutically acceptable excipient; packaged in a container with a label that describes use of the pharmaceutical composition. 
     
     
       23. A kit according to  claim 22 , wherein the pharmaceutical composition is packaged in a unit dose form. 
     
     
       24. A kit comprising a unit dose of a pharmaceutical composition according to  claim 1 , said kit comprising a first container comprising (a) a Factor VIII molecule; and (b) at least one water soluble polymer bound to said Factor VIII molecule, wherein said water soluble polymer is attached to the Factor VIII via one or more carbohydrate moieties located in the B domain of Factor VIII, wherein said molecule retains at least 50% of native FVIII activity, and a second container comprising a physiologically acceptable reconstitution solution for said conjugate in the first container, wherein said kit is packaged with a label that describes use of the pharmaceutical composition. 
     
     
       25. A kit according to  claim 24 , wherein the pharmaceutical composition is packaged in a unit dose form.